tbuckley/cabot-search huggingface dataset:
Abstract P2195: SARS-CoV-2 Variants Divergently Infect And Damage Cardiomyocytes
Authors: Bobo Mok, Maxwell Kwok, Hung Sing Li, Lowell Ling, Angel Lai, Paul Chan, Bryan Yan, Hong Lin Chen, Ellen Poon
Journal: Circulation Research (2023)

SARS-CoV-2 variants are known to exhibit different tropism and pathogenicity in respiratory cells, but their effect on cardiomyocytes (CMs) is unclear. Cardiac infection and damage induced by SARS-CoV-2 variants were examined using Golden Syrian hamsters and human induced pluripotent stem cell-derived (hiPSC-) CMs. Omicron BA.2 efficiently infected CMs in vitr o and in vivo , induced the most severe phenotype, and exhibited transcriptomic changes indicative of a more pro-inflammatory phenotype and increased cardiac dysfunction, compared to other variants. Increased infectivity of Omicron BA.2 is attributed to its ability to infect independently of TMPRSS2, which is absent in CMs. Our pilot study of patients infected with Omicron BA.2 in Hong Kong revealed significantly higher plasma troponin levels, consistent with severe myocardial injury. Here we reveal previously unknown differences in how different SARS-CoV-2 variants damage CMs, and that Omicron BA.2, which is generally considered mild, can induce severe CM damage in vitro , in vivo and in a subset of patients.
Abstract P1059: Decellularization Of Valved Porcine Xenografts For Use In Pediatric Right Ventricular Outflow Tract Replacement Surgery
Authors: Amy G Harris, Francesca Bartoli-Leonard, Dominga Iacobazzi, Michele Carraba, Giovanni Biglino, Massimo Caputo
Journal: Circulation Research (2023)

Introduction: The right ventricular outflow tract (RVOT) is pathologically affected in multiple congenital heart diseases, requiring surgical replacement within the first year of life. Shortage of small-sized homografts has driven xenograft use in pediatric cardiac surgery. However, these RVOT substitutes do not possess lifelong durability, eliciting high levels of inflammation and lacking growth potential. Xenograft limitations culminate in structural graft deterioration and rejection, resulting in invasive reoperations. Decellularization of donor tissue to eliminate cells and immunogenic antigens is a central method by which rejection risk can be diminished. Maintenance of the extracellular matrix (ECM) must be balanced with achievement of acellularity. Currently, no xenograft has proven optimal for pediatric RVOT surgical replacement; this work aims to produce a resilient and non-immunogenic decellularized porcine scaffold with translational capacity. Methods/Results: Research sought to optimize a combined chemical, enzymatic, and mechanical decellularization approach for RVOT conduits. The nine-day protocol utilizes a temperature-controlled bioreactor wherein EDTA, protease inhibitors, sodium dodecyl sulfate, and nucleases are circulated through the lumen of the submerged conduit. Maintenance of ECM structure, including elastin and collagen, in parallel with significant nuclei count reduction, removal of residual nucleic acids, and elimination of xenoantigens has been achieved. No significant difference was found between control and decellularized ventricular soluble collagen. All comparisons between samples processed directly after explant (fresh) and those decellularized following one freeze-thaw cycle (freeze-thawed) were not significantly different. Conclusion: Data indicate successful decellularization of valved conduits. Future work intends to seed mesenchymal stem cells onto the cell-free scaffold, improving biocompatibility and endowing the xenograft with growth potential. Decellularized RVOT xenografts provide a means to combat immunological mismatch between donor and patient, reducing the number of surgeries a child is exposed to and improving pediatric congenital surgery outcome.
Abstract P3076: Immune Checkpoint Regulator Impairs Left Ventricular Remodeling In A Hypertension-Fibrosis Model Of Cardiac Injury.
Authors: Jesus Jimenez, Pan Ma, Kory J Lavine
Journal: Circulation Research (2023)

Cancer treatments directly result in cardiovascular toxicity and may exacerbate typical age-related diseases such as hypertension and diabetes. Among cancer treatments contributing to survivorship, immunotherapies including immune checkpoint inhibitors (ICIs) are now widely utilized due to their groundbreaking efficacy. CD40 agonist (CD40ag) antibodies are immune checkpoint regulators that have emerged as promising candidates with remarkable efficacy across tumors including those thought to be resistant to established ICIs. To investigate CD40 signaling and its potential for adverse left ventricular remodeling in a hypertensive-fibrosis model of cardiac injury, C57BL/6J mice were pretreated with isotype or CD40ag antibodies 7 days prior to myocardial injury, then continued for a total of 35 days. Myocardial injury was induced by continuous infusion of angiotensin II and phenylephrine (AngII/PE) for 4 weeks. Masson’s trichrome stain revealed increased fibrosis and immunohistochemistry demonstrated increased immune cell abundance in heart sections. There was impaired cardiac function assessed by 2-D echocardiography in minimally sedated mice showing reduced ejection fraction and radial strain. Finally, CD40 agonism resulted in gene expression upregulation of multiple inflammatory cytokines and chemokines post-cardiac injury. These results reveal that pre-treatment with CD40 agonist prior to injury with AngII/PE infusion increased myocardial inflammation and impaired left ventricular remodeling.
Abstract P3096: Nitric Oxide Supplementation During Cardiac Arrest Induces Inflammation And Apoptosis In Rat Hearts
Authors: Abou Bakr M Salama, Daniele Linardi, Romel Mani, Maddalena M Tessari, Qinghui Ou, Bahaael Din El Sady, Ahmed M Shaker, Ahmed Abdulsaboor, Ahmad Gebreil, Riham Abouleisa, Giovanni Battista Luciani
Journal: Circulation Research (2023)

Background: Neuroprotection during cardiac and aortic arch surgeries has been a topic of extensive concern. Several protocols were developed in the trial to achieve the best neurological outcome; amongst them is nitric oxide (NO) supplementation through extracorporeal membrane oxygenation (ECMO). While NO was shown to influence the neurological outcome when supplemented during cardiac surgeries, a little is known about the cardiac effects. The current study addresses the molecular effects associated with NO supplementation on the myocardium. Methods: Male Sprague Dawley rats (n=20, 450-550gm) were anesthetized and cooled with cardiopulmonary bypass (CPB) to a core temperature of 19±1 o C and maintained in hypothermic cardiac arrest for 30 minutes. Then, rats were rewarmed to a target temperature of 35 o C for 60 min using CPB without (control group, CPB), or with NO 20 ppm supplementation (experimental group). By the end of experiment, hearts were harvested and analyzed. Results: RNA sequencing analysis showed NO supplementation results in the differential expression of more the genes related to lipid metabolism, oxidative phosphorylation, inflammatory signaling, proteasome formation, autophagy, calcium handling, smooth muscle relaxation, and apoptotic response compared to the control group. RT-PCR confirmed the RNA sequencing analysis. The experimental group showed a six-fold increase in inflammatory markers IL-1 and NF-κβ. The control group showed increased expression of α1 and β1 adrenergic receptors, NO treatment has led to normalization of expression and a three-fold increase in β2 receptors expression compared to shams. NO supplementation induced apoptosis in the myocardium, as indicated by a 50% increase in TUNEL-positive nuclei compared to the control group. Conclusions: NO supplementation during cardiac arrest boosts the inflammatory response in the myocardium and is associated with increased apoptosis.
Abstract P3035: Incidence, Complications And Outcomes Of Cerebral Venous Sinus Thrombosis In Patients With Acute Ischemic Stroke
Authors: Tilachan Parajuli, urza Niroula, Tanveer Mir, Shahzad Hassan, Waqas Qureshi, Abhishek Dutta, Shivani Mehta
Journal: Circulation Research (2023)

Introduction: Cerebral venous sinus thrombosis (CVST) is known to occur in patients with acute ischemic stroke. The incidence, complications and outcomes of CVST in acute ischemic stroke is unknown. We aimed to analyze a national database to evaluate the epidemiological characteristics of CVST in acute ischemic stroke patients. Method: We included weighted 624,046 adult patients (age ≥18 years) with acute ischemic stroke hospitalized in 2019 from the Nationwide Readmission Database (NRD). Baseline demographics, comorbidities, and outcomes were identified using International Classification of Diseases (ICD) codes. Results: Of the total, 1,156 (0.19%) stroke patients (mean age 50 ± 25) years and 55.9% females) were admitted with CVST. Of these, 144(12.4%) patients died in the CVST group and 41,372(6.64%) in the non CVST group (p &lt;0.001). Complications were noted to be higher in CVST group vs. non CVST; common complications that were significant included but not limited to; intracranial hemorrhage OR 6.11, CI (4.86-7.67), p &lt;0.001, resp failure OR 1.89, CI (1.47-2.44), p&lt;0.001, seizure OR 4.34, CI (2.74-6.87), cranial nerve palsy OR 8.15, CI (4.14-16.10), p&lt;0.001. The cost of index hospitalization and length of stay were higher for patients with CVST as well. Increased mortality was associated with diagnosis of CVST on presentation. Conclusion: In hospitalized acute ischemic stroke patients, cerebral venous sinus thrombosis was associated with increased risk of morbidity, mortality, increased length of stay and higher cost of hospitalization.
Abstract P2092: The Late Sodium Current Conditions Myocardial Repolarization And Relaxation Of The Aged Heart
Authors: Daniel O Cervantes, Emanuele Pizzo, Harshada Ketkar, Sreema P Parambath, Valeria Di Stefano, Kanwardeep Singh, Valentina Ripa, Anna Kouril, Peter J Mohler, Thomas J Hund, Jason Jacobson, Sudhir Jain, Marcello Rota
Journal: Circulation Research (2023)

Diastolic dysfunction and delayed ventricular repolarization are observed in the elderly, but whether these defects are intimately associated in the progressive manifestation of the aging myopathy remains to be determined. Interestingly, aging in experimental animals is coupled with increased late Na current (INaL) in cardiomyocytes, raising the possibility that INaL conditions the modality of electrical recovery and relaxation of the aged heart. Thus, male wild-type (WT) and genetically engineered mice with phosphomimetic (gain-of-function, GoF) or ablated (loss-of-function, LoF) mutations of the sodium channel Nav1.5 at Ser571, a phosphorylation site mediating INaL enhancement, were studied. With respect to WT mice at ~5 months of age (m), animals at ~18 m had 20% prolongation of the QT interval of the electrocardiogram and declined diastolic left ventricular filling, consisting of a 23% increase in isovolumic relaxation time (IVRT). These defects were reversed by INaL inhibition with GS967. Prolonged repolarization and impaired LV filling occurred prematurely in GoF mice (QT, +20%; IVRT, +8%, at ~5 m vs. WT), whereas age-related alterations were largely attenuated in LoF animals. At ~24 months, QT interval was not different in WT (66±4 ms) and GoF (70±5 ms) mice but was shorter in LoF (64±5 ms) animals, with respect to GoF. Similarly, IVRT was comparable in WT (22±2 ms) and GoF (22±2 ms) mice, but shorter in LoF (18±3 ms). In field stimulated cells, with respect to WT myocytes at ~5 m, WT cells at ~24 m had prolonged time to peak (+26%) and time to 50% relaxation (+44%). Delayed cell contraction/relaxation was apparent in GoF myocytes at ~5 m (+12% and +27%, with respect to WT), and further deteriorated with age. In contrast, LoF myocytes at ~5 m had contractile kinetics comparable to adult WT cells, and modest alterations occurred in cells at ~24 m. Importantly, inhibition of INaL was effective in correcting kinetics of cell mechanics in old WT mice and adult and old GoF animals but had no effect in LoF myocytes and WT cells at ~5 months. Collectively, these results document that the late Na+ current modulates the modality of myocyte relaxation, constituting the mechanism linking delayed ventricular repolarization and diastolic dysfunction in the elderly.
Abstract P2039: Alteration In Intercalated Disc Composition Impacts Cardiac Disease Progression
Authors: Janice Raabe, Konstantina Stathopoulou, Ilka Wittig, Jens Fielitz, Friederike Cuello
Journal: Circulation Research (2023)

Intercalated discs (IDs) connect cardiomyocytes electro-mechanically and ID dysfunction has been associated with the onset of cardiomyopathies and arrhythmias. The ID- and myofibril-associated protein nebulin-related anchoring protein (NRAP) was identified as a putative novel interaction partner of protein kinase D1 (PKD). PKD1 phosphorylates class IIa histone deacetylases (HDAC), leading to their cytosolic export. This causes a de-repression of HDAC-mediated inhibition of myocyte enhancer factor 2 (MEF2)-dependent fetal and pro-hypertrophic gene expression. We investigated the regulation of NRAP by PKD1 in the developing and diseased heart. PKD activity was modulated by using the pharmacological PKD inhibitor BPKDi and/or the pro-hypertrophic stimulus phenylephrine (PE) in neonatal rat ventricular myocytes (NRVMs) and -derived engineered heart tissues (EHTs). NRVMs were analyzed by immunofluorescence, western blotting and stable isotope labelling with amino acids in cell culture/mass spectrometry. The outcome was studied by exposure to an autophagy (bafilomycin A1) or a proteasomal (MG132) inhibitor or performing proteome analysis from hearts of MuRF-1/3 double-knock-out (DKO) mice. Exposure of NRVMs to PE enhanced myofibril maturation and NRAP translocation towards IDs while BPKDi led to perinuclear accumulation of NRAP and improved force in EHTs. Combined BPKDi and PE disturbed sarcomeric structure and prevented NRAP localization at the ID. Proteomic analysis revealed that BPKDi reduced NRAP abundance and enhanced the abundance of proteins involved in proteasomal, lysosomal and spliceosomal pathways. Inhibition of proteasomal protein degradation by MG132 increased NRAP levels, whereas NRAP was 42-fold more abundant in hearts of MuRF-1/3-DKO mice that show a protein surplus cardiomyopathy. In conclusion, PKD1 impacts NRAP expression and translocation, and negatively affects EHT force development while proteasomal inhibition increased NRAP abundance suggesting a regulation via the PKD1-HDAC-MEF2 axis. Further investigations aim to study pathways involving PKD1 to regulate myofibril assembly and maintenance in the development and progression of cardiac disease.
Abstract P3164: Podocalyxin, A Renal Podocyte Specific Protein Expression In Urine Exosome Acts As A Marker For Podocyte Injury During Sars-cov-2 Infection
Authors: Seshagiri Rao R Nandula, Sabyasachi Sen
Journal: Circulation Research (2023)

COVID-19 attributed to SARS-CoV2 infection has been associated with cardiovascular disease and diabetes. COVID-19 has been associated with endothelial cell dysfunction (ECD), which is manifested by entities such as microvascular thrombosis. ECD is quite common in type 2 diabetes mellitus (T2DM) where the renal podocyte dysfunction is often an early manifestation of microvascular complication. We explored whether presence of hyperglycemia predisposes to increased SARS-CoV2 infection and whether SARS-CoV2 infection puts an individual at a higher risk of developing cardio-metabolic complications such as kidney disease (DKD), with podocyte damage. We evaluated albuminuria and podocyte protein in urine derived exosomes in urine exosomes from SARS-CoV2 patients at 10 days, 6 months and 12 months post SARS-CoV2 infection. Methods: Blood and Urine samples from SARS-CoV2 patients post-acute phase of infection were procured. Peripheral blood mononuclear cells (PBMNCs) and urine exosomes were isolated and podocyte protein marker Podocalyxin (PODXL) was identified by western blot analysis. Results: Podocyte specific proteins were examined in urine exosomal fraction such as Podocalyxin and Nephrin (normalized to CD9 exosomal protein band intensities) levels increased significantly compared to urine samples from type-2 diabetes subjects with documented evidence of nephropathy. Podocalyxin levels were significantly high at 12month time point (p=0.001; n=9) and Nephrin levels were noted to be high at 10week (p=0.01; n=8), 6months (p=0.04; n=14 ) and 12Months (p=0.001; n=6) time points. Podocalyxin and Nephrin obtained from urine exosome samples collected from non diabetic subjects who never had COVID-19, showed little or barely detectable corresponding western blot bands. Interestingly, there was no significant differences noted on urine albumin: creatinine (UAR) ratios between the two groups, indicating that urine exosomal protein estimation may be an ideal biomarker to monitor podocyte damage in early phases of CKD. Conclusion: A persistent high levels of podocyte specific proteins was noted in urinary exosomes even at 12 months post acute Covid which may be secondary to long-standing podocyte inflammation leading to long term renal damage.
Abstract P3074: Alkylating Chemotherapy Reshapes The Immune Landscape Of The Heart
Authors: Ruijun He, Kory J Lavine
Journal: Circulation Research (2023)

Cancer survivors typically have concomitant cardiac comorbidities and were exposed to various chemotherapeutic agents. Despite extensive investigation, we still have a poor understanding of how chemotherapy agents impact cardiac physiology and disease pathogenesis. Alkylating agents, for instance, carboplatin, are the backbone of many chemotherapy regimens and have been applied in cancer treatment for decades. Cardiac macrophages are the primary component of the cardiac immune landscape and play essential roles in homeostasis and disease. My data implied that alkylating drugs induced cell death of the reparative cardiac macrophages (CCR2-) through activating apoptosis and necroptosis. However, the pro-inflammatory subset (CCR2+) is not decreased. Intriguingly, the reparative macrophages recovered four weeks after the carboplatin treatment through proliferation but showed different gene expression profiles. To investigate the functional influence of these reshaped macrophages in heart diseases, I established a hypertensive heart injury model and an ischemic heart injury model using carboplatin-exposed mice. Interestingly, these mice showed less fibrosis and smaller infarct size. Further, I used CD169-DTR mice that allow selective ablation of CCR2- macrophages. The attenuated remodeling phenotype is abolished without the reshaped CCR2- macrophages. Moreover, the type I interferon signaling in CCR2- macrophage is significantly upregulated in carboplatin-exposed mice upon hypertensive injury. Using neutralizing antibodies to block the IFN-I signaling, I found heart remodeling in carboplatin-treated mice became more advanced. Together, my study revealed that alkylating chemotherapy reshaped the immune landscape of the heart and mitigated adverse heart remodeling by activating the IFN-I signaling pathway in cardiac resident macrophages. These findings highlighted the role of CCR2- macrophages and the IFN-I signaling in the long-term cardiac effects of alkylating chemotherapy.
Abstract P3044: MicroRNA-127-3p Regulates Inflammatory Response In Injury-induced Vascular Remodeling And Cellular Senescence In Vascular Smooth Muscle Cells
Authors: Fiene Daniel, Katrin Kalies, Laura Hehl, Simon Guerlach, Jan Michael Köster, Solveig Boettcher, Kai Knoepp, Jochen Dutzmann, Daniel SEDDING
Journal: Circulation Research (2023)

The accumulation of senescent cells in the vasculature over the lifespan contributes to vascular remodeling due to the severely impaired functional properties of the cells. MicroRNAs (miRs) are promising therapeutical targets as they exhibit diverse and cell-specific functions and can be effectively targeted in vivo . Here, we propose that targeting miR-127-3p might be an attractive approach to enhance vascular healing and regeneration in a cell-specific manner, thereby limiting angioplasty-related complications. The initial screenings revealed miR-127-3p as a highly regulated miR in the process of restenosis 7 and 21 days (p&lt;0.001) after wire induced-injury and in aged (20 months) C57BL/6 mice (p&lt;0.05) in vivo . Additionally, replicative senescent vascular smooth muscle cells (SMC) and human coronary artery endothelial cells (EC) showed an enhanced expression of miR-127-3p (p&lt;0.05) compared to non-senescent cells. Functional effects of miR-127-3p on SMCs and ECs were assessed via transfection with either pre-miR or antagomir. Overexpression of miR-127-3p in vitro reduced SMC proliferation (p&lt;0.05) and altered migrational capacity. Downregulation of miR-127-3p resulted in a reverse effect. In contrast, cellular functions were not altered in ECs. We further identified potential targets with effects on inflammation and cellular aging by in silico analysis and verified them on mRNA and protein levels by qRT-PCR and western blot analysis. As a direct target of miR-127-3p, the senescence-associated gene CD44 was identified by TargetScan. Expression of CD44 was altered on mRNA and protein levels after overexpression respectively downregulation of miR-127-3p in SMCs. Further, p14arf and laminB1 were downregulated after overexpression of miR-127-3p in SMCs, contrary to the effect after downregulation of miR-127-3p with increased mRNA levels of p14arf and laminB1. In this study, we identified miR-127-3p as a contributing factor in SMC function during the development of cellular senescence and show that it might therewith also be a modulating factor in the process of restenosis. However, further studies need to be carried out to validate the potential of miR-127-3p as a therapeutic target in vascular aging and remodeling in vitro and in vivo .